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Purpose: The performance and validity of the microdialysis system in a time based calibration of the fluoride ion permeation inside the sheep eye following hydrofluoric acid burn was tested. The HF diffusion was measured in microdialysate samples after exposure to 0.15 ml of acid followed by neutralization with calcium gluconate formulations. The in vitro and in situ probe calibration and integrity test were performed. Methods: The stability of 10 BAS microdialysis probes was tested with 0.05 M HF by calculating the in vitro probe recovery. The effect of flow rate variation on recovery of fluoride ions was evaluated by performing an in vitro study for 0.01 M HF, with a constant sample volume of 40 μl at flow rates of 0.5, 1.0, 2.0 and 4.0 μl/min. The in situ probe calibration was performed on the aqueous and vitreous humor samples of the sheep eye, which could provide the baseline fluoride concentrations required for the calcium gluconate therapy. Two different formulations, solution (2.5% and 5.0% w/v calcium gluconate) and the 3% HPMC gel containing 2.5% w/v calcium gluconate were subjected to neutralization potential study. Results: The BAS probe was accurate and stable with a CV of 0.44-2.45%. A flow rate of 4.0 μl/min can be used for fluoride permeation study. The results showed that the MD probe is capable of dialyzing the acid without interacting with the probe. The in situ testing showed that the HPMC based calcium gluconate formulation showed longer residence time compared to the solution form. Conclusions: The microdialysis technique can be used to ascertain the fluoride ions penetrating in the HF-burnt eyes. The increase in amount of calcium ions available on eye surface, increases the free acid neutralization; the gel preparation being better than the solution of same concentration.
Microdialysis, Hydrofluoric acid diffusion, Fluoride permeation, in situ calibration of fluoride ions, Fluoride ion recovery, Drug Research and Discovery Process, Phases of Drug Development